To avoid the instability of an optical coefficient measurement using sliced tissue preparation, an attenuation coefficient measurement by puncturing an optical fiber in bulk tissue have been reported. We proposed a modified puncturing method to obtain an absorption coefficient and scattering coefficient by increasing optical information. We used the light intensity measurement through a single high-NA fiber puncturing into an optically thick bulk tissue varying detection parameters, the depth and field of view (FOV), at the tip. The light intensity measurement and ray trace calculation using the Monte Carlo method (inverse or normal) were employed. We constructed the measurement system which can change the FOV at the fiber tip inside the bulk tissue using the variable aperture outside the bulk tissue. A 200 μmΦ NA:0.5 fiber installed in a 21G needle was punctured down to surface of the bulk tissue to measure light intensity in the bulk tissue. Using homogeneous optical model solution, the accuracy about attenuation coefficients of the constructed experimental system was confirmed. The error in the attenuation coefficient was up to 1.4%. We demonstrated the optical coefficient measurement of porcine myocardium using the proposed method. To fit dependence of the measured attenuation coefficients on FOV, we decided that the absorption coefficient, the scattering coefficient, and anisotropic parameter were 0.2 mm-1, 10 mm-1, and 0.96, respectively. We proposed modified puncturing optical coefficient measurement varying depth and FOV. We demonstrated its usefulness on myocardium.
KEYWORDS: Tissue optics, Optical testing, Optical fibers, Signal attenuation, Monte Carlo methods, Ray tracing, Absorption, Scattering, Tissues, Geometrical optics
To avoid an instability of the optical coefficient measurement using sliced tissue preparation, we proposed the combination of light intensity measurement through an optical fiber puncturing into a bulk tissue varying field of view (FOV) and ray tracing calculation using Monte-Carlo method. The optical coefficients of myocardium such as absorption coefficient μa, scattering coefficient μs, and anisotropic parameter g are used in the myocardium optical propagation. Since optical coefficients obtained using thin sliced tissue could be instable because they are affected by dehydration and intracellular fluid effusion on the sample surface, variety of coefficients have been reported over individual optical differences of living samples.
The proposed method which combined the experiment using the bulk tissue with ray tracing calculation were performed. In this method, a 200 μmΦ high-NA silica fiber installed in a 21G needle was punctured up to the bottom of the myocardial bulk tissue over 3 cm in thickness to measure light intensity changing the fiber-tip depth and FOV. We found that the measured attenuation coefficients decreased as the FOV increased. The ray trace calculation represented the same FOV dependence in above mentioned experimental result. We think our particular fiber punctured measurement using bulk tissue varying FOV with Inverse Monte-Carlo method might be useful to obtain the optical coefficients to avoid sample preparation instabilities.
In order to study cardiomyocyte electrical conduction damage by a photosensitization reaction (PR) mostly comes from outside of the cells in a few minutes after the PR, we studied propagation delay of contact action potential with cardiomyocyte by the PR. To determine appropriate PR condition for tachyarrhythmia ablation, a precise electrophysiological experiment in vitro has been preferable. We measured the contact action potential using a microelectrode array system of which information may be correct than conventional Ca2+ measurement. We investigated the propagation delays of an evoked potential to evaluate the electrical conduction damage by the PR. Rat cardiomyocytes were cultivated for 5-7 days on a dish with which 64 electrodes were patterned, in an incubator controlled to 37°C, 5% CO2. The following conditions were used for the PR: 40 μg/ml talapordfin sodium and 290 mW/cm2, 40-78 J/cm2 for an irradiation. A 2D map was obtained to visualize the propagation delays of the evoked potential. The propagation speed, which was calculated based on the measured propagation delays, was decreased by about 30-50% on average of all electrodes after the PR. Therefore, we think 2D propagation delays measurement of the evoked potential with contact action potential measuring system might be available to evaluate the acute electrical conduction damage of cardiomyocyte by the PR.
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